Phospho-BIRC5 (Thr117) Antibody
Description
Introduction to Phospho-BIRC5 (Thr117) Antibody
Phospho-BIRC5 (Thr117) Antibody is a specialized reagent designed to detect phosphorylated survivin at the threonine 117 residue (Thr117). Survivin, encoded by the BIRC5 gene, is a multifunctional protein critical for mitosis regulation and apoptosis inhibition. The Thr117 phosphorylation site is a key post-translational modification (PTM) mediated by Aurora B kinase (AURKB), which modulates survivin’s interaction with chromosomal passenger complex (CPC) components and its subcellular localization during cell division .
Role of Thr117 Phosphorylation
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Regulation by Aurora B Kinase: Phosphorylation at Thr117 by AURKB disrupts survivin’s interaction with INCENP (a CPC component), preventing its localization to mitotic chromosomes .
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Mitotic Function: This modification inversely regulates survivin’s role in chromosome alignment, segregation, and cytokinesis .
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Anti-apoptotic Activity: Thr117 phosphorylation may destabilize survivin’s protective role against apoptosis during mitosis .
Subcellular Localization
Survivin dynamically localizes to:
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Centromeres (prophase to metaphase)
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Spindle midzone (anaphase)
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Midbody (telophase/cytokinesis) .
Phosphorylation at Thr117 reduces centromeric chromatin affinity, altering CPC localization .
Recommended Dilutions
| Application | Dilution Range |
|---|---|
| Western Blot | 1:500 – 1:2000 |
| IHC | 1:100 – 1:300 |
| IF/ICC | 1:200 – 1:1000 |
| ELISA | 1:5000 |
Research Use Cases
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Mitosis Studies: Investigate CPC dynamics during chromosome segregation .
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Cancer Research: Detect survivin overexpression in adenocarcinomas (lung, pancreas, colon) and lymphomas .
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Apoptosis Pathways: Study survivin’s dual roles in cell proliferation and caspase inhibition .
Post-Translational Modifications of Survivin
Validation and Quality Control
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Specificity: Recognizes endogenous survivin only when phosphorylated at Thr117 .
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Cross-Reactivity: Confirmed in human, mouse, and rat tissues; predicted reactivity in pig, bovine, and dog .
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Citations: RRID: AB_2834424 (Affinity Biosciences) ; STJ91292 (St John’s Labs) .
Clinical and Experimental Relevance
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Cancer Biomarker: Survivin is overexpressed in malignancies, making Thr117 phosphorylation a potential therapeutic target .
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Mitotic Dysregulation: Phospho-Thr117 status correlates with defects in cytokinesis and aneuploidy .
Limitations and Considerations
Product Specs
Target Background
- NF-kappaB activation may induce resistance to apoptosis through the expression of anti-apoptotic genes such as Survivin, which showed a progressive increase in expression throughout the esophageal metaplasia-dysplasia-adenocarcinoma sequence. PMID: 29473241
- This study demonstrates that ZIC1 plays a tumor suppressive role in breast cancer, by targeting Survivin, significantly downregulating its expression. PMID: 29956756
- This study observed that smoking patients had higher Survivin transcription and a remarkable spreading of Survivin isoforms, when studying Survivin expression in leukocytes of 144 female rheumatoid arthritis patients. PMID: 27915033
- Our findings identify Survivin as a target of HO-1 and a mediator of adipocyte-induced survival in the metastatic niche. PMID: 29311669
- High BIRC5 expression is associated with gefitinib resistance in lung cancer. PMID: 30106446
- miR-203 expression also inhibited primary tumor growth in ovaries and metastatic tumors in multiple peritoneal organs including liver and spleen. miR-203 inhibits ovarian tumor metastasis by targeting BIRC5/Survivin and attenuating the TGFbeta pathway. PMID: 30241553
- Survivin may be implicated in the bcl-2 and p53 pathways and therefore in the biology of PDAC. Its potential use as a survival predictor and therapeutic target represents a promising field. PMID: 30249893
- A study in hepatocellular carcinoma cell line elicited a new mechanism in which IGF-1 induced epithelial-mesenchymal transition through regulation of Survivin and a downstream pathway. PMID: 29989646
- In the present study, liposome-plasmid DNA encoding mutant SurvivinT34A could inhibit tumor growth of cervical cancer. This inhibition may be associated with an increase in the apoptosis rate of tumor cells and a reduction in angiogenesis. PMID: 29767242
- Simvastatin significantly inhibited the proliferation and invasion of SACC83 cells, induced apoptosis, and reduced the expression of Survivin, suggesting that simvastatin may be a novel target for salivary gland adenoid cystic carcinoma therapy. PMID: 29956779
- Survivin is significantly up-regulated in hepatocellular carcinoma (HCC) tissues and associated with tumor growth and lymph node metastasis. Clinical detection of Survivin levels combined with MRI examination might be beneficial for clinical diagnosis and treatment of HCC PMID: 30010107
- Our study identified the STAT3 rs1053004 C/C as a high-risk genotype in MA with lower Survivin and VEGF transcription levels in the peripheral blood. PMID: 30226700
- Overexpression of miR-485-5p suppresses breast cancer progression and enhances chemosensitivity. Further study demonstrated that miR-485-5p directly targeted the 3'-untranslated region of Survivin, and overexpression of Survivin overcomes the miR-485-5p induced effects on breast cancer. PMID: 29678577
- Survivin expression in gastric cancer cells is regulated by DEC1. PMID: 29204860
- High serum Survivin levels with GG genotype are associated with Brain Tumors. PMID: 30275230
- LNC473 could recruit deubiquitinase USP9X to inhibit the ubiquitination level of Survivin and then increase Survivin expression. PMID: 29605299
- Oct4 plays a vital role in the malignant progression of HCC cells through the Survivin/STAT3 signaling pathway. PMID: 29901157
- Our study results may suggest that high serum Survivin levels can show a 4 times increased risk of cancer in a subject with a high suspicion of cancer. Furthermore, Survivin level was not influenced with demographic characteristics of breast, gastric, colorectal, prostate, ovarian cancer, and glioblastome multiforme. PMID: 29893319
- These findings collectively suggest that the triple combination of Survivin knockdown with ABT-263 and trametinib treatment, may be a potential strategy for the treatment of KRAS-mutant lung adenocarcinoma. Furthermore, our findings indicate that the well-differentiated type of KRAS-mutant lung tumors depends, at least in part, on TTF1 for growth. PMID: 29658609
- Targeting the Cripto-1/TAK-1/NF-kappaB/Survivin pathway may be an effective approach to combat apoptosis resistance in cancer. PMID: 29807222
- Suggests that SIRT1 may serve as a predictor of poor prognosis in esophageal squamous cell carcinoma, and its mediated tumor-promoting role might be associated with the overexpression of EGFR protein in esophageal squamous cell carcinoma PMID: 29625788
- CSN5 directly bound Survivin and decreased its ubiquitination to enhance the protein stability of Survivin. PMID: 29596838
- The Survivin gene 3' UTR polymorphisms (rs17878624) show that the GG genotype provides substantial protection from non-small-cell lung carcinoma. PMID: 29631694
- Concomitant high expression of Survivin and VEGF-C is closely associated with LNM status of PTC patients, suggesting their cooperation in the metastatic process. PMID: 29578160
- In leukoplakia, the expression of Survivin associated with that of ki-67 reinforces the assumption that all these lesions are potentially malignant. PMID: 28346726
- This study showed for the first time that the suppression of rheumatoid arthritis fibroblast-like synoviocyte was mediated by phosphatase and tensin homolog involving Survivin silencing. PMID: 28337018
- ERCC1 expression may also inhibit esophageal squamous cell carcinoma cell apoptosis via regulating Survivin expression, and ERCC1 and Survivin overexpression are independent predictors of prognosis for ESCC patients who receive chemotherapy and/or radiotherapy PMID: 30075571
- let-7b targets PLK1 to inhibit hepatocellular carcinoma cell growth and induce their apoptosis by attenuating the PLK1-mediated Survivin phosphorylation PMID: 29913237
- Overexpression of Survivin in glioma cells induces chromosomal instability PMID: 29282022
- High BIRC5 expression is associated with ovarian cancer. PMID: 29795564
- In the third trimester of pregnancy, parameters of Timp-1 and Survivin - anti-apoptotic substances concentration were similar in maternal and cord blood in both artery and vein. We found no increased activity of selected antiapoptotic factors. PMID: 28509321
- Results from a study in non-small-cell lung cancer cells showed that SphK2 plays a critical role in doxorubicin-induced resistance by regulating key anti-apoptotic gene, Survivin. PMID: 28950390
- Survivin overexpression plays a key role in the chemoresistance of ovarian CSCs. PMID: 30061219
- Survivin may play an important role in the occurrence and development of laryngeal carcinoma, and its high expression is related to the poor prognosis of patients with laryngeal cancer. (Meta-analysis) PMID: 29270761
- Treatment of DLD1 cells with tamoxifen, beta-estradiol, or a combination of these two drugs, inhibited cell viability and migration, promoted cell apoptosis, and reduced the mRNA and protein expression levels of Survivin in a dose and time-dependent manner. These results provide novel experimental basis for hormonal adjuvant therapy for the treatment of colorectal cancers PMID: 28849238
- Studies indicated that high Survivin expression in renal cell carcinoma (RCC) was associated with poor overall survival [Review]. PMID: 27411378
- Nuclear accumulation of Survivin is associated with a proliferative phenotype and was shown to be a worse prognostic marker in breast ductal carcinoma. PMID: 29517199
- Knockdown of BIRC5, a member of the inhibitor of apoptosis protein family, using either lentiviral vector-based CRISPR/Cas9 nickase gene editing or inhibition of Survivin using the small-molecule inhibitor YM155, results in the suppression of epithelial to mesenchymal transition in retinal pigment epithelial cells. PMID: 29522718
- By downregulation of Sp1 and Survivin at the late phase of treatment. PMID: 28713892
- The expressions of Survivin and STAT2 are up-regulated in skin lesions of PV patients, and their mRNA expressions are positively correlated. PMID: 29089085
- Findings suggest that lysosome-associated transmembrane protein 4B (LAPTM4B), vascular endothelial growth factor (VEGF), and nuclear Survivin expression are significantly correlated in breast cancer, which may be predictive of prognosis as well as effective therapeutic targets for anticancer therapies. PMID: 28476037
- HIF-2alpha dictates the resistance of human pancreatic cancer cells to TRAIL under normoxic and hypoxic conditions and transcriptionally regulates Survivin expression. PMID: 28476028
- Survivin may have a role in recurrence in rectal cancer patients treated with surgery and postoperative concurrent chemo-radiation therapy PMID: 27391438
- Results identified BIRC5 to be significantly upregulated in the lung squamous cell carcinoma tissues of smoking patients and may play an important role in diagnosis and prognosis. PMID: 28949095
- After cancer cell fusion, some fused cells avoid the apoptotic crisis partly owing to Survivin, and continue to proliferate, a process that contributes to human cancer progression. PMID: 28193315
- High Survivin expression is associated with lung cancer and colorectal cancer. PMID: 27602754
- Data suggest that Ki-67 index and Survivin may be useful biomarkers for rectal cancer with preoperative chemoradiotherapy. PMID: 29491110
- Inhibition of apoptosis targeting Survivin might represent an effective strategy for both obesity and cancer therapy. PMID: 28518147
- FAT10 promotes tumor proliferation by directly stabilizing Survivin protein in breast cancer cells. PMID: 27806337
- Nuclear Survivin is a prognostic marker for the progression of oral squamous cell carcinomas. PMID: 28384094
Q&A
What is Phospho-BIRC5 (Thr117) Antibody and what are its specifications?
Phospho-BIRC5 (Thr117) Antibody is a rabbit polyclonal antibody specifically designed to detect Survivin protein only when phosphorylated at the Threonine 117 position . This antibody is produced through affinity purification via sequential chromatography on phospho- and non-phospho-peptide affinity columns to ensure high specificity . Key specifications include:
| Specification | Details |
|---|---|
| Target Protein | Phospho-Survivin (Thr117) |
| Clonality | Polyclonal |
| Host Species | Rabbit |
| Reactivity | Human, Mouse, Rat |
| Predicted Cross-Reactivity | Pig, Bovine, Horse, Sheep, Dog |
| Molecular Weight | 16 kDa |
| Applications | WB, IHC, IF/ICC |
| UniProt Accession | O15392 |
The antibody is typically stored in phosphate-buffered saline (pH 7.4) containing 150mM NaCl, 0.02% sodium azide, and 50% glycerol, and remains stable for approximately 12 months when stored at -20°C .
How does phosphorylation at Thr117 affect Survivin function?
Survivin undergoes several post-translational modifications, with phosphorylation being particularly important for regulating its varied cellular functions . While phosphorylation at Thr34 is well-documented for its role in anti-apoptotic activity, Thr117 phosphorylation represents a distinct regulatory mechanism .
Phosphorylation at different sites provides survivin with diverse molecular functions:
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Thr34 phosphorylation (regulated by CDK1/P34cdc2 cyclin B1) enhances cytoprotective effects and is necessary for anti-apoptotic activity
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Thr117 phosphorylation modulates survivin's role in cell cycle regulation and potentially its interactions with other chromosome passenger complex components
Unlike other Inhibitor of Apoptosis Proteins (IAPs), survivin contains only one Baculoviral IAP Repeat (BIR) domain, highlighting the significance of its phosphorylation sites in determining functional specificity .
What tissue distribution patterns are observed for Survivin expression?
Survivin exhibits a highly regulated expression pattern that varies significantly between normal and pathological conditions:
Normal Tissues:
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Expressed predominantly in fetal kidney and liver
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Lower expression in fetal lung and brain
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Present in cochlea including the organ of Corti, lateral wall, interdental cells of the Limbus
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Found in Schwann cells and cells of the cochlear nerve and spiral ganglions
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Notably absent in cells of the inner and outer sulcus or the Reissner's membrane
Pathological Conditions:
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Abundantly expressed in various adenocarcinomas (lung, pancreas, colon, breast, and prostate)
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Highly expressed in high-grade lymphomas
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Present in renal cell carcinoma cell lines
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Elevated expression in PBMCs of patients with Behcet's disease
This distinctive expression pattern makes survivin and its phosphorylated forms potential biomarkers for various pathological conditions.
What methodological approaches are recommended for detecting phosphorylated survivin in different sample types?
Detection of phosphorylated survivin requires careful methodological consideration depending on sample type and research objectives:
For PBMC Samples:
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Cell harvesting followed by protein extraction using appropriate lysis buffers
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Western blot analysis using anti-phosphorylated-survivin antibodies (specific to the phosphorylation site of interest)
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Comparison with total survivin levels using pan-survivin antibodies
For Plasma Samples:
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Collection of plasma with appropriate anticoagulants
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Protein precipitation or direct analysis depending on concentration
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Western blot analysis for qualitative assessment
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Enzyme-linked immunosorbent assay (ELISA) for quantitative measurement
Recommended dilutions for various applications:
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Western Blot: 1:500-1:2000
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Immunohistochemistry: 1:50-1:200
How can researchers quantify BIRC5 mRNA expression alongside protein phosphorylation studies?
A comprehensive analysis of survivin biology often requires examination at both mRNA and protein levels. For BIRC5 mRNA quantification:
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Extract total RNA from PBMCs using high-quality RNA isolation kits
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Synthesize cDNA using reverse transcription
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Perform quantitative real-time PCR using specific primers:
This approach allows correlation between mRNA expression and phosphorylation status to understand both transcriptional and post-translational regulation.
What are the differences in phosphorylated survivin expression between plasma and PBMCs in disease states?
Research on Behcet's disease has revealed intriguing differences in phosphorylated survivin expression between cellular and circulating compartments:
PBMCs:
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Increased expression of phosphorylated survivin in Behcet's disease patients compared to healthy controls
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Particularly elevated phosphorylation at Thr34 in patients with active disease
Plasma:
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Downregulation of phosphorylated survivin levels in Behcet's disease patients compared to healthy individuals
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Total survivin levels in plasma show no significant differences between patients and controls (p>0.05)
This differential expression suggests compartment-specific regulation of survivin phosphorylation and potential translocation mechanisms that may contribute to disease pathogenesis.
How does phosphorylated survivin contribute to autoimmune pathogenesis?
Survivin plays multiple roles in immune cell function, with phosphorylation status influencing its contribution to autoimmune pathogenesis:
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Apoptosis Dysregulation: Defective apoptosis is a fundamental problem in autoimmune diseases, and survivin acts as an anti-apoptotic protein that can inhibit caspase 3 function
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Cell Cycle Disruption: Survivin interferes during the G2/M phase of mitosis in deregulated cell cycle checkpoints to promote abnormal cell survival
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Immune Cell Development: Survivin is critical for proliferation, maturation, homeostasis, and differentiation of effector and memory lymphocytes
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Gene Expression Changes: BIRC5 gene expression is increased in Behcet's disease patients compared to healthy controls (p<0.05)
The increased expression of phosphorylated survivin in PBMCs of autoimmune patients suggests it could serve as a potential biomarker for disease activity and therapy monitoring .
What are the known splice variants of BIRC5 and how might they affect antibody detection?
The BIRC5 gene encodes several spliced variants with different amino acid compositions:
| Variant | Length (amino acids) | Potential Effect on Antibody Detection |
|---|---|---|
| Wild type | 142 | Standard detection target |
| ΔEx3 | 137 | May affect epitope accessibility |
| 2B | 165 | Additional amino acids could alter conformation |
| 3B | 120 | Shortened protein may lack Thr117 site |
| 2α | 74 | Likely lacks Thr117 phosphorylation site |
| 3α | 78 | Likely lacks Thr117 phosphorylation site |
Researchers should be aware that alternative splicing may affect the presence or accessibility of the Thr117 phosphorylation site . Validation experiments using positive and negative controls are essential when working with different cell types or tissues that might express varying splice variants.
